Abstract
Field surveys were conducted in Greek olive orchards from 2017 to 2020 to collect information on the sanitary status of the trees. Using a high-throughput sequencing approach, viral sequences were identified in total RNA extracts from several trees and assembled to reconstruct the complete genomes of two isolates of a new viral species of the genus Tepovirus (Betaflexiviridae), for which the name olive virus T (OlVT) is proposed. A reverse transcription–polymerase chain reaction assay was developed which detected OlVT in samples collected in olive growing regions in Central and Northern Greece, showing a virus prevalence of 4.4% in the olive trees screened. Sequences of amplified fragments from the movement–coat protein region of OlVT isolates varied from 75.64% to 99.35%. Three olive varieties (Koroneiki, Arbequina and Frantoio) were infected with OlVT via grafting to confirm a graft-transmissible agent, but virus infections remained latent. In addition, cucumber mosaic virus, olive leaf yellowing-associated virus and cherry leaf roll virus were identified.
Highlights
Olives (Olea europaea L.) have been cultivated in the Mediterranean area since prehistoric times
Among the seven known olive viruses tested for, only three (CMV, cherry leaf roll virus (CLRV), olive leaf yellowing associated virus (OLYaV)) were detected: cucumber mosaic virus (CMV) was detected in five trees sampled in Agrinio, Attica, Argolida and Korinthia; OLYaV was detected in two trees, in Arkadia (Koutroufa) in the Peloponnese and Kilkis (Evropos) in Macedonia; and CLRV was detected in one tree in each of two areas (Megara and Paiania) in Attica (Figure 4)
Graft transmission experiments unambiguously demonstrated its identity as a novel transmissible agent, for which the name olive virus T (OlVT) is proposed
Summary
Olives (Olea europaea L.) have been cultivated in the Mediterranean area since prehistoric times. In Greece, graft-transmissible, virus-like diseases of olive trees have been reported [4]; upto-date information on the occurrence of viruses in Greek olive orchards is scarce. Pairwise analysis at the whole genome level revealed 81.0% nt sequence identity obfe tween GR168 and GR170, and 74.5% and 74.8% nt identity, respectively, to the closest tepovirus, PrVT (KF700262). Among the seven known olive viruses tested for, only three (CMV, CLRV, OLYaV) were detected: CMV was detected in five trees sampled in Agrinio, Attica, Argolida and Korinthia; OLYaV was detected in two trees, in Arkadia (Koutroufa) in the Peloponnese and Kilkis (Evropos) in Macedonia; and CLRV was detected in one tree in each of two areas (Megara and Paiania) in Attica (Figure 4). CLRV 3’ UTR 2.5.3U. sCiLngRVth3e’ CULTRRV specific primers, we amplified an identical 341 bp long fragment from tUhsein3g’ UthTeRCrLegRiVonspfoercitfhice ptwriomGerrse,ewk eolaimvepilsifoileadteasn(GidRe1n8t6icOaLl 3a4n1dbGpRlo2n52gOfrLa;gGmeenn-t Bfarnokmatchcees3s’ioUnTsRMrWeg8i1o4n91fo3ratnhde MtwWo8G1r4e9e1k4,orleisvpeeicstoivlaetleys).(GThRe1s8e6sOeLquaenndceGs Rsh2o52wOeLd;oGnelyn9B1.a9n%k saicmceislasrioitnyswMitWh 8tw14o91o3thaenrdidMeWnti8c1a4l9G14re, erkesopleivcteiviseolyla).teTshaelsreeasedqyupeuncbeliscslyhoawvaeidlaobnlely (M91K.99%36s2i3m5,ilMarKit9y3w62i3th6).twHoigohtehresreiqdueenntciceaslimGrileaerkityol(i9v3e.8i4s%ol)awteassaolrbesaedrvyepduwbiltichlya Pavlaanitlaagbole m(MajoKr 9(K36F273759,2M05K) 9a3n6d23a6n).AHctiignhideira scehqinueennscise(sKimF7i7la9r2i0ty2)(i9s3o.8la4t%e,)bwotahsforobmserNveedwwZietahlaanPdl.antago major (KF779205) and an Actinidia chinensis (KF779202) isolate, both from New Zealand
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